RESUMO
Interspecies transmissions of viruses between animals and humans may result in unpredictable pathogenic potential and new transmissible diseases. This mechanism has recently been exemplified by the discovery of new pathogenic viruses, such as the novel severe acute respiratory syndrome corona virus-2 (SARS-CoV-2) pandemic, Middle-East respiratory syndrome-coronavirus epidemic in Saudi Arabia, and the deadly outbreak of Ebola in West Africa. The. SARS-CoV-2 causes coronavirus disease-19 (COVID-19), which is having a massive global impact in terms of economic disruption, and, above all, human health. The disease is characterized by dry cough, fever, fatigue, myalgia, and dyspnea. Other symptoms include headache, sore throat, rhinorrhea, and gastrointestinal disorders. Pneumonia appears to be the most common and severe manifestation of the infection. Currently, there is no vaccine or specific drug for COVID-19. Further, the development of new antiviral requires a considerable length of time and effort for drug design and validation. Therefore, repurposing the use of natural compounds can provide alternatives and can support therapy against COVID-19. In this review, we comprehensively discuss the prophylactic and supportive therapeutic role of probiotics for the management of COVID-19. In addition, the unique role of probiotics to modulate the gut microbe and assert gut homeostasis and production of interferon as an antiviral mechanism is described. Further, the regulatory role of probiotics on gut-lung axis and mucosal immune system for the potential antiviral mechanisms is reviewed and discussed.Key points⢠Gut microbiota role in antiviral diseases⢠Factors influencing the antiviral mechanism⢠Probiotics and Covid-19.
Assuntos
Betacoronavirus/fisiologia , Infecções por Coronavirus/terapia , Pneumonia Viral/terapia , Probióticos/uso terapêutico , Animais , COVID-19 , Infecções por Coronavirus/prevenção & controle , Infecções por Coronavirus/transmissão , Trato Gastrointestinal/microbiologia , Humanos , Imunidade nas Mucosas , Pulmão/imunologia , Pulmão/microbiologia , Pulmão/virologia , Pandemias/prevenção & controle , Pneumonia Viral/prevenção & controle , Pneumonia Viral/transmissão , Probióticos/metabolismo , Infecções Respiratórias/microbiologia , SARS-CoV-2 , Viroses/prevenção & controle , Viroses/terapia , Viroses/transmissão , Vitamina D/fisiologia , Zinco/metabolismoRESUMO
Beta-hexosaminidase (Hex) is the major lysosomal enzyme associated with the event of fertilization. In this study, we have analyzed the distribution of Hex in the testis and the epididymis of the lizard, Eutropis carinata by a polyclonal antibody of ß-hexosaminidase isoform (Hex A). Presence of Hex in the epididymis was performed by Western blotting. The result reveals that Hex A is present in the epididymal epithelium, lumen as well as spermatozoa. The anatomical distribution of Hex was studied by immunohistochemical localization. The study reveals that Hex is intensely stained in the epithelium of anterior and middle regions of the epididymis, whereas, posterior epididymal epithelium shows moderate staining. In addition, seminiferous epithelium of the testis shows staining for Hex. But lumen of the testis did not show any reaction for Hex. Further, immunohistochemical localization of Hex on the spermatozoa from the testis and different regions of the epididymis revealed that the Hex from the testis did not show any staining; the epididymal epithelium is moderately localized in the spermatozoa of the anterior region and gradually increases in the intensity in the spermatozoa of the posterior region of the epididymis. This indicates that the Hex is released from the epididymal epithelium and binds to the spermatozoa, and in the lumen, it gradually increases from anterior to the posterior region of the epididymis. The result also suggests that Hex A bound to the epididymal spermatozoa originates from the epididymis and not from the testis. The regional difference in the expression of Hex in the epididymis of the lizard, E. carinata, indicates the possible site of secretion of this enzyme.
Assuntos
Genitália Masculina/enzimologia , Lagartos/metabolismo , beta-N-Acetil-Hexosaminidases/metabolismo , Animais , Western Blotting , Genitália Masculina/citologia , Imuno-Histoquímica , MasculinoRESUMO
Type 2 diabetes mellitus (T2DM) is a chronic metabolic disorder characterized by hyperglycemia and insulin resistance. Gut microbiota (GM) are specific groups of microbes colonized in the gastrointestinal (GI) tract. They profoundly influence health, disease protection, and associated with metabolic activities, and play a vital role in the production of functional metabolites from dietary substances. Dysbiosis of GM has been linked to the onset of T2DM and can be altered to attain eubiosis by intervention with various nutritional bioactive compounds such as polyphenols, prebiotics, and probiotics. This review presents an overview of the evidence and underlying mechanisms by which bioactive compounds modulate the GM for the prevention and management of T2DM.
Assuntos
Diabetes Mellitus Tipo 2 , Microbioma Gastrointestinal , Probióticos , Diabetes Mellitus Tipo 2/tratamento farmacológico , Diabetes Mellitus Tipo 2/prevenção & controle , Disbiose , Humanos , Prebióticos , Probióticos/uso terapêuticoRESUMO
Endurance refers to the ability of skeletal muscles to perform continuously withstanding the hardships of exercise. Endurance exercises have three phases: pre-, during-, and post-workout phase. The nutritional requirements that drive these phases vary on intensity, type of workout, individual's body composition, training, weather conditions, etc. Generally, the pre-workout phase requires glycogen synthesis and spare glycogen breakdown. While workout phase, requires rapid absorption of exogenous glucose, insulin release to transport glucose into muscle cells, replenish the loss of electrolytes, promote fluid retention, etc. However, post-workout phase requires quick amino acid absorption, muscle protein synthesis, repair of damaged muscle fibres and tendon, ameliorate inflammation, oxidative stress, etc. Therefore, nutritional sources that can help these metabolic requirements is recommended. In this review, various dietary interventions including timing and amount of nutrient consumption that can promote the above metabolic requirements that in turn support in improving the endurance potential in athletes are discussed.HIGHLIGHTSReview article describes nutritional requirements of endurance exercises.It also describes nutritional interventions to enhance the endurance potential in athletes.
Assuntos
Suplementos Nutricionais , Resistência Física , Atletas , Glucose , Glicogênio/metabolismo , Humanos , Músculo Esquelético/metabolismo , Resistência Física/fisiologiaRESUMO
Since the discovery of NAD-dependent deacetylases, sirtuins, it has been recognized that maintaining intracellular levels of NAD is crucial for the management of stress response of cells. Here we show that agonist-induced cardiac hypertrophy is associated with loss of intracellular levels of NAD, but not exercise-induced physiologic hypertrophy. Exogenous addition of NAD was capable of maintaining intracellular levels of NAD and blocking the agonist-induced cardiac hypertrophic response in vitro as well as in vivo. NAD treatment blocked the activation of pro-hypertrophic Akt1 signaling, and augmented the activity of anti-hypertrophic LKB1-AMPK signaling in the heart, which prevented subsequent induction of mTOR-mediated protein synthesis. By using gene knock-out and transgenic mouse models of SIRT3 and SIRT1, we showed that the anti-hypertrophic effects of exogenous NAD are mediated through activation of SIRT3, but not SIRT1. SIRT3 deacetylates and activates LKB1, thus augmenting the activity of the LKB1-AMPK pathway. These results reveal a novel role of NAD as an inhibitor of cardiac hypertrophic signaling, and suggest that prevention of NAD depletion may be critical in the treatment of cardiac hypertrophy and heart failure.
Assuntos
Proteínas Quinases Ativadas por AMP/metabolismo , NAD/química , Proteínas Serina-Treonina Quinases/metabolismo , Sirtuína 3/metabolismo , Quinases Proteína-Quinases Ativadas por AMP , Animais , Cardiomegalia/patologia , Insuficiência Cardíaca , Hipertrofia , Camundongos , Camundongos Transgênicos , Ligação Proteica , Ratos , Espécies Reativas de OxigênioRESUMO
Newcastle disease (ND) is highly contagious, economically important viral disease affecting most of avian species worldwide. Newcastle disease virus (NDV) has single stranded negative sense RNA genome which encodes for six structural and two non-structural proteins. Envelope glycoproteins i.e. hemagglutinin-neuraminidase (HN) and the fusion (F), elicit protective immune response. In this study, HN and F genes of velogenic (virulent) strain were amplified and cloned at multiple cloning sites A and B, respectively into pIRES bicistronic vector for use as bivalent DNA vaccine against ND. The recombinant plasmid was characterized for its orientation by restriction enzyme digestion and PCR. Expression of HN and F genes was assessed in transfected Vero cells at RNA level using RT-PCR in total RNA as well as protein level using IFAT, IPT and western blot using NDV specific antiserum. All these experiments confirmed that HN and F genes cloned in recombinant pIRES.nd.hn.f are functionally active. The recombinant construct is being evaluated as DNA vaccine against ND.
Assuntos
Clonagem Molecular , Proteína HN/genética , Vírus da Doença de Newcastle/genética , Vacinas de DNA/genética , Proteínas Virais de Fusão/genética , Animais , Chlorocebus aethiops , DNA Viral/genética , Vírus da Doença de Newcastle/imunologia , Proteínas Recombinantes de Fusão/genética , Transfecção , Vacinas de DNA/imunologia , Células Vero , Proteínas Virais de Fusão/imunologiaRESUMO
Preeclampsia is a pregnancy-related multisystem disorder, frequently encountered pregnancy-related medical complications next to gestational diabetes mellitus. It is the onset of hypertension during pregnancy. The preeclampsia can be of two types, placental or maternal preeclampsia. Among these two types former, i.e., placental preeclampsia is more severe than the latter. According to the recent survey by National Health Portal of India, the incidence of preeclampsia is about 8-10 % among pregnant women. Though our understanding of preeclampsia has improved in recent years, the development and interpretation of the clinical tests remain difficult for preeclampsia. Hence, we have made an attempt to understand the pathophysiology, associated conditions/consequences, treatment and management/prevention of the condition in this review.
Assuntos
Pré-Eclâmpsia/prevenção & controle , Pré-Eclâmpsia/fisiopatologia , Autoanticorpos/sangue , Citocinas/metabolismo , Parto Obstétrico , Feminino , Humanos , Hipóxia/fisiopatologia , Neovascularização Patológica/fisiopatologia , Óxido Nítrico/biossíntese , Estresse Oxidativo/fisiologia , Placenta/fisiopatologia , Insuficiência Placentária/fisiopatologia , Gravidez , Receptor Tipo 1 de Angiotensina/imunologiaRESUMO
The COVID-19 is an acute and contagious disease characterized by pneumonia and ARDS. The disease is caused by SARS-CoV-2, which belongs to the family of Coronaviridae along with MERS-CoV and SARS-CoV-1. The virus has the positive-sense RNA as its genome encoding for ~26 proteins that work together for the virus survival, replication, and spread in the host. The virus gets transmitted through the contact of aerosol droplets from infected persons. The pathogenesis of COVID-19 is highly complex and involves suppression of host antiviral and innate immune response, induction of oxidative stress followed by hyper inflammation described as the "cytokine storm," causing the acute lung injury, tissue fibrosis, and pneumonia. Currently, several vaccines and drugs are being evaluated for their efficacy, safety, and for determination of doses for COVID-19 and this requires considerable time for their validation. Therefore, exploring the repurposing of natural compounds may provide alternatives against COVID-19. Several nutraceuticals have a proven ability of immune-boosting, antiviral, antioxidant, anti-inflammatory effects. These include Zn, vitamin D, vitamin C, curcumin, cinnamaldehyde, probiotics, selenium, lactoferrin, quercetin, etc. Grouping some of these phytonutrients in the right combination in the form of a food supplement may help to boost the immune system, prevent virus spread, preclude the disease progression to severe stage, and further suppress the hyper inflammation providing both prophylactic and therapeutic support against COVID-19.
Assuntos
Antivirais/uso terapêutico , Infecções por Coronavirus/dietoterapia , Infecções por Coronavirus/tratamento farmacológico , Reposicionamento de Medicamentos/métodos , Compostos Fitoquímicos/uso terapêutico , Pneumonia Viral/dietoterapia , Pneumonia Viral/tratamento farmacológico , Anti-Inflamatórios/uso terapêutico , Antioxidantes/uso terapêutico , Betacoronavirus/efeitos dos fármacos , COVID-19 , Infecções por Coronavirus/patologia , Síndrome da Liberação de Citocina/dietoterapia , Síndrome da Liberação de Citocina/tratamento farmacológico , Síndrome da Liberação de Citocina/patologia , Citocinas/sangue , Suplementos Nutricionais , Humanos , Inflamação/tratamento farmacológico , Estresse Oxidativo/fisiologia , Pandemias , Pneumonia Viral/patologia , Probióticos/uso terapêutico , SARS-CoV-2RESUMO
The velogenic Newcastle disease virus (NDV) causes highly infectious and economically significant Newcastle disease (ND) in birds of various species. In cell culture NDV induces cytopathic effect (CPE) characterized by rounding, vacuolation, syncytia formation and cell death. Aside from cell to cell fusion caused by the F and HN glycoprotein of the virus molecular events leading to cell death are not known. In the current study, NDV-infected Vero cells, at 48 h p.i., showed nuclear condensation, cytoplasm blebbing, DNA fragmentation, and phosphatidylserine translocation to the cell surface. In addition, virus-infected cells demonstrated decreased DNA content and an increased Bax to Bcl-2 ratio, p53 level and caspase 3, 8, 9 expression compared to mock-infected cells. Based on these results, it was concluded that CPE in NDV-infected cells was caused by to the induction of apoptosis with the involvement of p53 and the Bax, dependent apoptotic pathways.
Assuntos
Apoptose , Doença de Newcastle/fisiopatologia , Doença de Newcastle/virologia , Vírus da Doença de Newcastle/fisiologia , Animais , Caspases/genética , Caspases/metabolismo , Chlorocebus aethiops , Efeito Citopatogênico Viral , Fragmentação do DNA , Expressão Gênica , Doença de Newcastle/genética , Doença de Newcastle/metabolismo , Proteínas Proto-Oncogênicas c-bcl-2/genética , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Células VeroRESUMO
Cancer is a major cause of deaths in humans. Though there has been significant progress in cancer therapy, the limited efficacy and toxicities of current chemo- and radiotherapies have provided an impetus for the search of new therapeutics. A therapeutic approach, which uses viruses for the treatment of cancer termed, oncolytic virotherapy has recently emerged. Newcastle disease virus (NDV) is one such virus with an inherent oncolytic property. NDV causes a highly infectious disease in poultry worldwide. In humans it is reported to have oncolytic and immuno-stimulatory effects. It specifically replicates in tumour cells while sparing normal cells and cause oncolysis. For many years different strains of the NDV have been investigated for treatment of various human cancers. Recent advances in reverse genetics provided investigators the tools to produce recombinant NDV with improved oncolytic property.
Assuntos
Neoplasias/terapia , Vírus da Doença de Newcastle/fisiologia , Terapia Viral Oncolítica/métodos , Animais , Apoptose , Humanos , Neoplasias/patologia , Vírus da Doença de Newcastle/genética , Vírus Oncolíticos/genética , Vírus Oncolíticos/fisiologiaRESUMO
Polycystic ovary syndrome (PCOS) is one of the major endocrine disorders in women. PCOS is a disorder with alterations in the structure as well as functions of female reproductive organs and is also associated with metabolic disorders. Studies on humans have limitations due to ethical issues, hence animal models are used to understand the different aspects of PCOS. Animal models of PCOS aids in studying various facts beginning from etiology to the treatment, hence, several animal models have been developed. Despite of the establishment of several models and a number of studies on PCOS, lacunae exist. This may be due to the failure in selecting a suitable animal model, as all animal models may not exhibit all the key features of the human PCOS condition or may exhibit traits similar to other diseased conditions in addition to the PCOS which should be excluded. Therefore, in this review, we have discussed the different animal models, features they exhibit, their merits and limitations which may aid in the selection of the relevant animal model of PCOS based upon the investigation's focus. In addition, a few nonmammalian models as an alternative to mammalian models have also been discussed which is to be validated further.
Assuntos
Modelos Animais de Doenças , Sistema Endócrino/fisiopatologia , Síndrome do Ovário Policístico/etiologia , Síndrome do Ovário Policístico/patologia , Animais , Feminino , HumanosRESUMO
Newcastle disease virus causes (NDV) apoptotic death of infected cells. In the present study, the stimulus that provoked the induction of apoptosis in infected cells was examined. Vero cells infected with NDV developed apoptosis as characterized by DNA fragmentation and decreased DNA content. In presence of ammonium chloride, infected cells did not show reduced DNA content indicating the requirement of virus entry for the induction of apoptosis. UV-inactivated NDV did not induce apoptosis in cells suggesting the need of virus replication. Although cycloheximide blocked NDV-induced apoptosis, actinomycin-D did not, suggesting that de-novo viral protein synthesis was critical for the induction of apoptosis. In addition, activation of caspases was also detected by flowcytometry, indirect fluorescent and colorimetric assays. Based on the results, it was concluded that NDV-induced apoptosis in Vero cells required virus replication, de-novo protein synthesis and caspase activation.
Assuntos
Apoptose , Caspases/metabolismo , Vírus da Doença de Newcastle/patogenicidade , Proteínas Virais/metabolismo , Replicação Viral , Animais , Chlorocebus aethiops , Fragmentação do DNA , Ativação Enzimática , Vírus da Doença de Newcastle/fisiologia , Células VeroRESUMO
The mechanism of postovulatory follicle (POF) regression in birds is still poorly understood. In the current study, expression of IL-1beta, IL-6, GM-CSF, IFN-gamma, IL-2, IL-4, IL-13, chCXCLi2, chCCLi2, chCCLi4, chCCLi7, IL-10 and TGF-beta2 mRNAs was estimated in regressing POF by semi-quantitative RT-PCR. In addition, the changes in immune cell population, histological and apoptotic changes were also studied in regressing POF. The expression of cytokines (IL-1beta, IL-6, IL-10 and TGF-beta2) and chemokines (chCXCLi2, chCCLi2, chCCLi4 and chCCLi7) was upregulated in POFs, suggesting a role for these molecules in tissue regression. The histological findings suggested a significant infiltration of immune cells, especially heterophils, lymphocytes and macrophages, into the regressing POF. The flow cytometry analysis of lymphocyte subpopulations revealed that CD3(+), CD4(+), CD8(+) and Bu-1(+) lymphocytes were significantly increased during this regression. The significant up-regulation of chemokines might have attracted the immune cells during POF regression. The percentage of apoptotic cells was significantly increased during the regression of POF. The up-regulation of IL-1beta, IL-6, IL-10 and TGF-beta2 and down-regulation of GM-CSF might have induced apoptosis during the POF regression. However, expression of IFN-gamma, IL-2, IL-4 and IL-13 was not significantly altered during POF regression. In conclusion, cytokines appear to play an important role in the regression of POF in chicken. Furthermore, the regression of chicken POF seems to be an inflammatory event similar to luteolysis of the mammalian corpus luteum.
Assuntos
Quimiocinas/genética , Galinhas/genética , Citocinas/genética , Folículo Ovariano/metabolismo , Animais , Apoptose/genética , Apoptose/fisiologia , Galinhas/fisiologia , Feminino , Expressão Gênica , Interferon gama/genética , Interleucinas/genética , Subpopulações de Linfócitos/citologia , Subpopulações de Linfócitos/metabolismo , Folículo Ovariano/citologia , Ovário/citologia , Ovário/metabolismo , Ovulação/genética , Ovulação/fisiologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fator de Crescimento Transformador beta2/genéticaRESUMO
The present study was aimed at developing the polymerase chain reaction (PCR) assay for detection of canine adenoviruses from faecal or urine samples. Urine or faecal samples were treated with chloroform or activated charcoal to eliminate the PCR inhibitory substances and the total DNA was extracted. The PCR was optimized using common set of primers to amplify 508 bp or 1,030 bp DNA sequence within E3 gene of canine adenovirus-1 (CAV-1) and canine adenovirus-2 (CAV-2), respectively. The PCR assay could detect up to 0.016 TCID(50) viruses from CAV-1 infected MDCK cell culture fluid, 1.6 TCID(50) viruses from faeces and 16 TCID(50) viruses from urine. In addition, the PCR assay was validated using clinical samples. Based on the results, it is concluded that, the present PCR assay can be used for rapid detection of canine adenoviral infections.
Assuntos
Infecções por Adenoviridae/diagnóstico , Adenovirus Caninos/isolamento & purificação , Doenças do Cão/diagnóstico , Fezes/virologia , Reação em Cadeia da Polimerase/métodos , Urina/virologia , Infecções por Adenoviridae/virologia , Proteínas E3 de Adenovirus/genética , Adenovirus Caninos/genética , Animais , Linhagem Celular , Primers do DNA/genética , DNA Viral/isolamento & purificação , Doenças do Cão/virologia , Cães , Sensibilidade e EspecificidadeRESUMO
1. The immune responses induced by recombinant plasmids containing Newcastle disease virus (NDV) F (pVAX.nd.f) or HN (pcDNA.nd.hn) genes separately or in combination in bi-cistronic (pIRES.nd.hn.f) constructs were evaluated in maternal antibody-positive commercial chicks. 2. Immunofluorescence and immunoperoxidase tests demonstrated the expression of both F and HN proteins in Vero cells. Real-time PCR analysis revealed the expression of HN and/or F genes in muscle, peripheral blood mononuclear cells (PBMC), spleen and liver after immunisation. 3. Chicks inoculated intramuscularly thrice (two booster doses) with pVAX.nd.f and pcDNA.nd.hn did not develop detectable haemagglutination inhibiting (HI) antibodies. In contrast, an increase in a NDV-specific cell-mediated immune response was demonstrated. 4. After challenge with virulent NDV, chicks immunised with the recombinant plasmids as well as those in control groups succumbed to Newcastle disease. 5. Based on these results, it is concluded that DNA vaccines containing HN and/or F genes fail to protect commercial chicks, possibly due to interference from maternal antibodies.
Assuntos
Galinhas/imunologia , Hemaglutininas Virais/genética , Vírus da Doença de Newcastle/genética , Vacinas de DNA/imunologia , Proteínas Virais de Fusão/genética , Animais , Anticorpos Antivirais/sangue , Galinhas/virologia , Chlorocebus aethiops , Feminino , Imunofluorescência , Expressão Gênica , Vetores Genéticos , Imunidade Celular , Técnicas Imunoenzimáticas , Doença de Newcastle/imunologia , Doença de Newcastle/prevenção & controle , Plasmídeos/genética , Proteínas Recombinantes/genética , Células VeroRESUMO
In the long term, diabetes profoundly affects multiple organs, such as the kidney, heart, brain, liver, and eyes. The gradual loss of function in these vital organs contributes to mortality. Nonetheless, the effects of diabetes on the lung tissue are not well understood. Clinical and experimental data from our studies revealed that diabetes induces inflammatory and fibrotic changes in the lung. These changes were mediated by TGF-ß-activated epithelial-to-mesenchymal transition (EMT) signaling pathways. Our studies also found that glucose restriction promoted mesenchymal-to-epithelial transition (MET) and substantially reversed inflammatory and fibrotic changes, suggesting that diabetes-induced EMT was mediated in part by the effects of hyperglycemia. Additionally, the persistent exposure of diabetic cells to high glucose concentrations (25 mM) promoted the upregulation of caveolin-1, N-cadherin, SIRT3, SIRT7 and lactate levels, suggesting that long-term diabetes may promote cell proliferation. Taken together, our results demonstrate for the first time that diabetes induces fibrotic changes in the lung via TGF-ß1-activated EMT pathways and that elevated SMAD7 partially protects the lung during the initial stages of diabetes. These findings have implications for the management of patients with diabetes.
Assuntos
Diabetes Mellitus/genética , Fibrose Pulmonar/genética , Transdução de Sinais/genética , Fator de Crescimento Transformador beta1/genética , Animais , Diabetes Mellitus/metabolismo , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/metabolismo , Transição Epitelial-Mesenquimal/efeitos dos fármacos , Transição Epitelial-Mesenquimal/genética , Regulação da Expressão Gênica/efeitos dos fármacos , Glucose/farmacologia , Humanos , Rim/efeitos dos fármacos , Rim/metabolismo , Rim/patologia , Pulmão/efeitos dos fármacos , Pulmão/metabolismo , Pulmão/patologia , Masculino , Fibrose Pulmonar/metabolismo , Ratos Wistar , Transdução de Sinais/efeitos dos fármacos , Proteína Smad7/genética , Proteína Smad7/metabolismo , Fator de Crescimento Transformador beta1/metabolismo , Fator de Crescimento Transformador beta1/farmacologiaRESUMO
Abnormal activation of insulin-like growth factor (IGF)-Akt signaling is implicated in the development of various diseases, including heart failure. However, the molecular mechanisms that regulate activation of this signaling pathway are not completely understood. Here we show that sirtuin 6 (SIRT6), a nuclear histone deacetylase, functions at the level of chromatin to directly attenuate IGF-Akt signaling. SIRT6-deficient mice developed cardiac hypertrophy and heart failure, whereas SIRT6 transgenic mice were protected from hypertrophic stimuli, indicating that SIRT6 acts as a negative regulator of cardiac hypertrophy. SIRT6-deficient mouse hearts showed hyperactivation of IGF signaling-related genes and their downstream targets. Mechanistically, SIRT6 binds to and suppresses the promoter of IGF signaling-related genes by interacting with c-Jun and deacetylating histone 3 at Lys9 (H3K9). We also found reduced SIRT6 expression in human failing hearts. These findings disclose a new link between SIRT6 and IGF-Akt signaling and implicate SIRT6 in the development of cardiac hypertrophy and failure.
Assuntos
Cardiomegalia , Insuficiência Cardíaca , Proteínas Quinases JNK Ativadas por Mitógeno , Proteína Oncogênica v-akt , Sirtuínas , Acetilação , Animais , Cardiomegalia/genética , Cardiomegalia/metabolismo , Cromatina/genética , Cromatina/metabolismo , Proteínas de Ligação a DNA/genética , Proteínas de Ligação a DNA/metabolismo , Insuficiência Cardíaca/genética , Insuficiência Cardíaca/metabolismo , Histona Desmetilases/metabolismo , Fator de Crescimento Insulin-Like I/genética , Fator de Crescimento Insulin-Like I/metabolismo , Camundongos , Camundongos Transgênicos , Proteína Oncogênica v-akt/genética , Proteína Oncogênica v-akt/metabolismo , Regiões Promotoras Genéticas , Transdução de Sinais , Sirtuínas/deficiência , Sirtuínas/genética , Sirtuínas/metabolismoRESUMO
Classical swine fever (CSF) is an economically important Office International des Epizooties (OIE) list A disease of swine characterized by high fever and multiple haemmorhages. The E2 glycoprotein of CSFV is immunogenic and induces neutralizing antibodies against CSFV. In the present study, complete coding region of the E2 gene from Indian virulent field isolate (Mathura) was amplified by reverse transcription-polymerase chain reaction (RT-PCR) and subsequently cloned into a mammalian expression vector; pcDNA3.1(+) at BamHI and XbaI site. The recombinant plasmid; pcDNA.E2.CSFV. was confirmed by restriction enzyme digestion. The pcDNA.E2.CSFV. transfected Vero cell expressed E2 protein which was confirmed by western blotting, immunoperoxidase and indirect immunofluorescent tests. Additionally, flow cytometry analysis also confirmed that 15% of transfected Vero cells expressed the E2 glycoprotein compared to mock or vector alone transfected cells. Further study is under way to evaluate recombinant pcDNA.E2.CSFV. Mathura clone as DNA vaccine against CSFV.
RESUMO
Newcastle disease virus (NDV) causes economically significant Newcastle disease (ND) in almost all birds worldwide. Previous studies have shown that NDV induces caspase dependent apoptotic pathways in infected cells. In the present study, time course induction of apoptotic pathways in Vero cells is described. In NDV-infected cells, caspase-8 activity, percentage of cells showing TRAIL expression was higher at 24h p.i. (post-infection) compared to 48 h p.i. In contrast, caspase-9 activity, efflux of cytochrome c, loss of mitochondrial membrane potential was higher at 48 h compared to 24h p.i. The caspase-3 activity was high both times. Based on these results, it was concluded that at 24h p.i., NDV induces apoptosis through extrinsic apoptotic pathway while at 48 h p.i. predominantly through intrinsic apoptotic pathway.
Assuntos
Apoptose , Vírus da Doença de Newcastle/patogenicidade , Animais , Caspase 8/metabolismo , Chlorocebus aethiops , Citocromos c/análise , Citoplasma/química , Perfilação da Expressão Gênica , Potencial da Membrana Mitocondrial/fisiologia , Ligante Indutor de Apoptose Relacionado a TNF/metabolismo , Fatores de Tempo , Células VeroRESUMO
Newcastle disease virus (NDV), an avian paramyxovirus, induces apoptosis in chicken embryo fibroblast (CEF) cells. In the present investigation, the ability of haemagglutinin-neuraminidase (HN) protein of NDV to cause apoptosis in CEF cells was examined. The results revealed that cells expressing the HN protein demonstrated decreased DNA content, phosphatidylserine exposure and increased cytoplasmic vacuolation. Up-regulation of caspase-1, -9, -8, -3, loss of mitochondrial transmembrane potential and an increase in oxidative stress were also observed in cells expressing the HN protein. Based on the above results it can be concluded that HN protein of NDV causes apoptosis in CEF cells.